Legumes establish symbiosis with rhizobia by forming a symbiotic interface that enables cross-kingdom exchanges of signaling molecules and nutrients. However, how host organelles interact with symbiosomes at the symbiotic interface remains elusive during rhizobia endosymbiosis. Here, symbiotic cells are reconstructed using 3D scanning electron microscopy (SEM) and uncover that the host endoplasmic reticulum (ER) undergoes dynamic expansion to gradually enwrap symbiosomes, facilitating their compartmentalization and endosymbiosis. Consistently, altering ER lamellar expansion by overexpressing MtRTNLBs, the reticulons responsible for ER tubulation, impairs rhizobia accommodation and symbiosome development. Intriguingly, unfolded protein response (UPR)-marker genes, bZIP60 and IRE1A/B, show continuously activated expression during nodule development, and the two UPR-deficient mutants, ire1b, and bzip60, exhibit compromised ER biogenesis and defective symbiosome development. Collectively, the findings underpin ER expansion and UPR activation as two key events in rhizobia accommodation and reveal an intrinsic coupling of ER morphology with proper UPR during root nodule symbiosis.